How to Make DTT Solution?
To prepare a DTT solution, precisely dissolve 150 g of DL-dithiothreitol in approximately 800 mL of distilled water, then carefully bring the total volume to 1 L with additional distilled water. This potent stock solution is then dispensed into small aliquots for long-term storage, ensuring its stability and effectiveness.
Dithiothreitol (DTT) is a widely used reducing agent in biochemistry and molecular biology, essential for preventing oxidation of protein sulfhydryl groups and for reducing disulfide bonds. Creating a concentrated stock solution like this ensures a ready supply for various laboratory applications.
Required Materials
To prepare this specific DTT stock solution, you will need the following:
- DTT (DL-dithiothreitol, anhydrous): 150 g (molecular weight = 154.25 g/mol)
- Distilled Water: Approximately 1 L
- Suitable Container: A volumetric flask or graduated cylinder capable of holding 1 L
- Spatula or Weigh Boat: For handling DTT
- Weighing Scale: Capable of accurately measuring 150 g
- Parafilm or Stopper: For sealing the container
- 1-mL Aliquot Tubes: For dispensing the final solution
- Aluminum Foil: For wrapping aliquots to protect from light
- Freezer: Capable of maintaining -20°C
Step-by-Step Preparation
Follow these steps carefully to ensure accurate and effective DTT solution preparation:
- Initial Water Addition: Begin by adding approximately 800 mL of distilled water to a suitable container, such as a 1-L volumetric flask or a large beaker.
- Add DTT: Carefully weigh out 150 g of DL-dithiothreitol. Add the measured DTT to the distilled water in your container.
- Dissolve DTT: Stir or swirl the solution gently to ensure the DTT completely dissolves. DTT dissolves readily in water.
- Adjust Final Volume: Once the DTT is fully dissolved, add additional distilled water until the total volume of the solution precisely reaches 1 L. Ensure the bottom of the meniscus aligns with the 1-L mark if using a volumetric flask.
- Mix Thoroughly: Cap the container and invert it several times to ensure thorough mixing of the solution.
Storage and Longevity
Proper storage is crucial for maintaining the stability and efficacy of DTT solutions, especially concentrated stock solutions like this one.
- Aliquot Dispensing: Dispense the freshly prepared 1-L DTT solution into 1-mL aliquots. This practice minimizes the number of freeze-thaw cycles for the entire stock, which can degrade the DTT over time.
- Dark Storage: Wrap each 1-mL aliquot in aluminum foil or store them in amber tubes. DTT is sensitive to light, and protecting it from light exposure helps maintain its stability.
- Temperature: Store the aliquots in a freezer at -20°C. At this temperature, the DTT solution can be stored indefinitely, providing a long-lasting supply for your laboratory needs.
Quick Reference: DTT Solution Preparation
For a concise overview, refer to the table below detailing the components for this specific DTT solution:
| Component | Quantity | Notes |
|---|---|---|
| DTT (DL-dithiothreitol) | 150 g | Anhydrous, m.w. = 154.25 g/mol |
| Distilled Water (initial) | 800 mL | For initial dissolution |
| Distilled Water (final) | Add up to 1 L | To reach final volume |
| Final Volume | 1 L | Target volume of the stock solution |
| Aliquot Size | 1 mL | For storage to preserve long-term stability |
| Storage Temperature | -20°C | Store indefinitely |
| Storage Condition | Dark (wrapped) | Protect from light |
